Aim. Cultivation of Entamoeba histolytica is usually performed for diagnostic and experimental purposes. In this study, it is aimed to determine the effects of different physical conditions on the growth of E. histolytica Bangladesh strain in Robinson and Locke Egg Serum (LES) media. Methods. Parallel to regular passages at 35.5ºC in Robinson and LES media, incubation conditions for the 37.5ºC and %5 CO2 incubator is investigated. Additionally, the growth rates were further investigated in vitro at 6.5 and 7.5 pH values. Results. It was observed that at 37.5ºC trophozoites were able to survive but reproduction rate was decreased. Five percent CO2 incubator and standard bacteriological incubator did not yield difference in terms of parasite reproduction. No living trophozoite was detected in pH changed media on fourth day after passage. Conclusions. Among the changes in the cultivation of E. histolytica, pH has a crucial effect on parasite growth and %5 CO2 incubator can be used instead of standard incubator for incubation.